1. Field of the Invention
The present invention relates to a method for highly sensitive nucleic acid detection using a nanopore and a non-specific nucleic acid-binding agent.
2. Description of the Related Art
Various methods of detecting a target biomolecule in a sample have been reported. A method using nanopores is widely used in a highly sensitive DNA detecting system, which is an imitation of a bio-pore system, and is capable of sequencing bases in nucleic acids.
A method of characterizing individual polymer molecules based on monomer-interface interactions is disclosed in U.S. Pat. No. 6,362,002. In this method, a single-stranded nucleic acid polymer can pass through a channel in an interface between two pools, but a double-stranded nucleic acid polymer cannot pass through the channel, and can thus be detected.
A method of determining the presence of double-stranded nucleic acids in a sample is disclosed in U.S. Pat. No. 6,428,959. Double-stranded nucleic acid can be detected by translocating nucleic acids in a sample through a nanopore, monitoring a current amplitude through the nanopore during the translocation, and measuring the duration of a transient blockade of current. However, U.S. Pat. No. 6,428,959 does not disclose a method of detecting nucleic acids using a non-specific nucleic acid-binding agent to increase a current amplitude.
A method for the characterization of nucleic acid molecules is disclosed in U.S. Patent Publication No. 2003/0104428. A maximum change in a signal can be achieved by changing specific local areas using proteins specific to nucleotide sequences. However, U.S. Patent publication No. 2003/0104428 relates to the detecting of DNA having a specific sequence using nanopore, and does not disclose the detecting of nucleic acids using a non-specific nucleic acid-binding agent to increase a current amplitude.
The inventors of the present invention discovered that a nucleic acid can be accurately detected by increasing a current amplitude change through nanopores using a non-specific nucleic acid binding agent, regardless of specific sequences.